Chloroform-free permeabilization for improved detection of β-galactosidase activity in Vibrio cholerae

Charlotte Toulouse; Claudia C Häse; Julia Steuber

doi:10.1016/j.mimet.2017.03.011

Chloroform-free permeabilization for improved detection of β-galactosidase activity in Vibrio cholerae

J Microbiol Methods. 2017 Jun:137:1-2. doi: 10.1016/j.mimet.2017.03.011. Epub 2017 Mar 21.

Authors

Charlotte Toulouse¹, Claudia C Häse², Julia Steuber³

Affiliations

¹ Institute of Microbiology, University of Hohenheim (Stuttgart), 70599 Stuttgart, Germany.
² Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
³ Institute of Microbiology, University of Hohenheim (Stuttgart), 70599 Stuttgart, Germany. Electronic address: julia.steuber@uni-hohenheim.de.

PMID: 28336461
DOI: 10.1016/j.mimet.2017.03.011

Abstract

LacZ (β-galactosidase) is used to monitor the transcription of genes in reporter strains carrying the lacZ gene under the control of a promotor of interest. This protocol for LacZ activity determinations in Vibrio cholerae following detergent lysis results in 2.5-fold increase of LacZ activities compared to lysis with chloroform.

Keywords: Bacterial cell lysis; LacZ; Vibrio cholerae; β-Galactosidase.

MeSH terms

Bacterial Proteins / genetics
Cell Membrane / metabolism
Chloroform / pharmacology*
Detergents / pharmacology
Genes, Reporter
Lac Operon
Nitrophenols / metabolism
Promoter Regions, Genetic
Protease Inhibitors / pharmacology
Transcription, Genetic
Vibrio cholerae / drug effects*
Vibrio cholerae / enzymology*
Vibrio cholerae / genetics
beta-Galactosidase / genetics
beta-Galactosidase / metabolism*

Substances

Bacterial Proteins
Detergents
Nitrophenols
Protease Inhibitors
Chloroform
beta-Galactosidase